Detection of the Milk Proteins by RP-HPLC (in English)

Abstract

In this study the bovine κ-, α- and β-caseins, α-lactalbumin and β-lactoglobulin by HPLC-UV using an RP column was detected by developed a simple and rapid method. Gradient elution was carried out at a 1 ml/min flow rate, and a temperature of 25 oC, using a mixture of two solvents. Solvent A was acetonitrile: water: trifluoroacetic acid (100:900:1) and Solvent B was acetonitrile:water:trifluoroacetic acid (900:100:1). The effluent flow was monitored by a UV detector at 220 nm. Different chromatographic profiles were obtained for bovine milk proteins which was prepared with different methods for RP-HPLC. It was suggested that using of Solvent A and Solvent B during the sample preparation by addition into the sample before injection at given ratio was allowed the separation more clear and effective than the other sample preparation methods. By the current sample preparation technique and elution practice which was occurred in this study, it was achieved detection of bovine milk proteins was being simple, rapid and sensitive provide.

Keywords

• Milk, proteins, caseins, whey proteins, RP-HPLC.　　

Süt Proteinlerinin RP-HPLC ile Saptanması (İngilizce)

Abstract

Bu çalışmada, κ-, α- and β-kazein ile α-laktalbumin and β-laktoglobulin, bir RP kolonunun kullanıldığı HPLC-UV ile basit ve hızlı bir yöntem geliştirilerek saptanmıştır. Gradient elüsyonu, 1 ml/dk akış hızında ve 25 oC sıcaklıkta iki çözücü karışımı kullanılarak yürütülmüştür. A çözücüsü asetonitril: su:trifloroasetik asit (100:900:1) ve B çözücüsü asetonitril: su: trifloroasetik asit (100:900:1) karışımlarından oluşmaktadır. Akış, bir UV dedektörü kullanılarak 220 nm’de kaydedilmiştir. Farklı yöntemler kullanılarak RP-HPLC analizi için hazırlanan örneklerde, süt proteinlerine ait farklı kromatografik profiller elde edilmiştir. Diğer örnek hazırlama yöntemlerine kıyasla, enjeksiyondan önce belirtilen oranlarda A ve B çözücülerinin kullanıldığı örnek hazırlama yöntemi ile separasyonun daha keskin ve efektif olduğu ortaya konulmuştur. Bu çalışmada ortaya konulan örnek hazırlama tekniği ve elusyon pratiği ile, süt proteinlerinin analizi basit, hızlı ve duyarlı bir şekilde gerçekleştirilmiştir.

Keywords

• Süt, proteinler, kazeinler, serum proteinleri, RP-HPLC.      　　　

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